Computational protocol: A multigene family encoding surface glycoproteins inTrypanosoma congolense

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Protocol publication

[…] Plastic-adherent trypanosomes grown in culture were scrapped, collected by centrifugation (500 g, 5 min, RT) and washed in gPBS.For whole-cell and « shaving » experiments, trypanosomes were fixed with paraformaldehyde (PFA, 3%) as described elsewhere .For cytoskeleton-extracted cell experiments, parasites were extracted with Pipes buffer (100 mM Pipes pH 6.9, 1 mM MgCl2, 0.05% NP40) for 3 min, washed and fixed with PFA (3%).Slides were incubated with mouse anti-TcoRep (diluted 1:100) or mouse anti-beta tubulin (diluted 1:200) followed by Alexa Fluor 596-conjugated goat anti-mouse secondary antibody (diluted 1:100) (Invitrogen). The nucleus and the kinetoplast were stained with DAPI (10 µg/mL). Cells were viewed using a Zeiss Axio Imager Z1 fluorescent microscope; images were captured using the MetaMorph® software (Molecular Devices) and processed using ImageJ software. […]

Pipeline specifications

Software tools MetaMorph, ImageJ
Application Microscopic phenotype analysis
Organisms Homo sapiens
Diseases Trypanosomiasis
Chemicals Amino Acids